WO2013057567A1 - Biomarqueurs servant à détecter des types, niveaux et stades du cancer du sein chez l'être humain - Google Patents

Biomarqueurs servant à détecter des types, niveaux et stades du cancer du sein chez l'être humain Download PDF

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WO2013057567A1
WO2013057567A1 PCT/IB2012/002090 IB2012002090W WO2013057567A1 WO 2013057567 A1 WO2013057567 A1 WO 2013057567A1 IB 2012002090 W IB2012002090 W IB 2012002090W WO 2013057567 A1 WO2013057567 A1 WO 2013057567A1
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breast cancer
biomarkers
seq
grade
panel
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PCT/IB2012/002090
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Lekha DINESH KUMAR
Vinod Kumar VERMA
Rekha A. NAIR
Jem PRABHAKAR
Jayasree KATTOOR
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Council Of Scientific And Industrial Research (C.S.I.R.)
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Priority to CA2852384A priority Critical patent/CA2852384C/fr
Priority to US14/352,804 priority patent/US10208353B2/en
Priority to AU2012324531A priority patent/AU2012324531B2/en
Priority to GB1408696.1A priority patent/GB2509682B/en
Publication of WO2013057567A1 publication Critical patent/WO2013057567A1/fr
Priority to AU2018202963A priority patent/AU2018202963B2/en
Priority to US16/219,572 priority patent/US11136628B2/en

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    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA

Definitions

  • the present invention relates to a panel of biomarkers useful for detection of types, grades and stages of human breast cancer.
  • the present invention particularly relates to the development of these identified biomarkers as a miRNA chip for the early and accurate diagnosis of human breast cancer.
  • This patent application highlights the novelty in the utility of these miRNAs, that they could be used as a diagnostic kit (miRNA chip) for early and accurate detection of breast cancer grades, stages and subtypes.
  • Tissue characterization by pathologists for ER, PR and HER 2/Neu status and axillary lymph node status are the most important prognostic factors and 90% of those patients without nodal involvement have no further breast cancers detected in their lifetime.
  • Axillary nodal status is of major importance from a therapeutic and prognostic point of view.
  • Chemotherapeutic drugs currently used are also not specific to breast cancer. Therefore it is imperative to find novel biomarkers for early and accurate diagnosis and prognosis in breast cancer sparing the majority of patients from undergoing an axillary dissection.
  • Such molecular signatures can also lead to good prognosis and help develop novel targeted treatments.
  • such an approach can accurately identify subgroups of patients who will really benefit from cytotoxic chemotherapy with its debilitating side effects.
  • the diagnosis of breast tumor starts with the screening techniques to confirm whether a lump is present or not.
  • the noninvasive examination techniques existing are mammography, ultrasound or MR imaging which determine the presence of any tumors and also detect tumor size, invasion etc.
  • FNAC Fine Needle Aspiration and Cytology
  • Additional testing may include genetic screening that test for the status of hormones like ER, PR, and genes like HER2/neu etc. Chemotherapy is currently used for all cases of Infiltrating duct carcinomas of breast.
  • Mammography and ultrasound may identify a potential area of concern.
  • MRI imaging requires injection of a dye, the side effects of which are not yet proven.
  • Fine Needle Aspiration Cytology (FNAC) is not always as reliable as surgical biopsies in producing a conclusive diagnosis.
  • Immunohistochemical analysis of ER, PR, HER2/neu, BRCA and PTEN requires lot of time to arrive at any final conclusion of disease progression.
  • the available diagnostic methods present in the market are not up to the expectation that one can diagnose the early stages of disease and therapeutic measures can be optimized to completely prevent and cure the tumor at right time.
  • These above mentioned imaging tools are not sensitive methods to detect early molecular changes occurring in the cell during initiation of the cancer.
  • Tissue embedding, sectioning, staining are all cumbersome procedures and time consuming. Moreover, staging could be determined only after getting the final histopathology report and extensive metastatic workup. No existing technologies are there for more accurate staging of the disease for identifying suitable patient sub groups to tailor systemic treatment.*There are no proper fast and accurate molecular diagnostic tool for pathologists till now for accurate staging and grading. As far as current chemotherapy regimens are concerned, no targeted therapy is currently used.
  • Biomarkers constitute the most important field in cancer diagnosis. Cancer biomarkers are especially useful for early detection or diagnosis of the disease. Biomarkers can be used to screen patients, for classifying the different stages or grades of cancers and to predict prognosis and resistance to therapy. A tumour marker can be produced by tumour itself or by the body as a result of the disease. These biomolecules are quite often produced in abnormally large numbers in the cancerous
  • MicroRNAs are small RNAs of 22-25 nucleotides in length with a major role in gene regulation. Since they are highly conserved between the genomes of related species and show a characteristic evolutionary divergence, computational analysis of miRNAs would augment the experimental analysis to identify those which are involved in the regulation of common genes and pathways leading to the development of cancer. Recently, oncomiRs, special classes of non-coding microRNAs are found to be associated with a large number of cancers. Consequently impaired miRNA expression is implicated in various tumours. This class of novel non-coding RNAs or microRNAs is expected to eventually identify previously unappreciated tumour suppressors and oncogenes and also address many questions about the origin, development and progression of breast cancer.
  • miRNAs are upregulated: hsa-miR-141, hsa-miR-200b,
  • miRNAs are Down regulated:
  • hsa-miR- 125b- l has-miR125b- 2
  • has-miR-145 has-miR-145
  • hsa -miR-21 has-miR-155
  • OncomiRs are a special class of non-coding microRNAs found to be associated with a large number of cancers. Consequently impaired miRNA expression is implicated in various tumours.
  • Various in vitro and in vivo studies have implicated an active role of microRNAs in breast cancer. Many reports on microRNAs indicated their role in cell proliferation and apoptosis growth and migration (1&2) suggesting that deregulation of these microRNA could lead to proliferative diseases like cancer. Also studies have shown that microRNA cluster mapped to the hotspot areas of the genome that are prone for cancer mutations (3 &4). Their expression patterns show a general trend of down regulation in human cancer samples (5) indicating that most of them function as tumour suppressors.
  • microRNA miR 206 was found to inhibit the function of estrogen receptor gene ESR1. Later, it was found to be targeted by a set of microRNAs like miR 18a, miR 18b, miR193b and miR 302c (6 & 7). CyclinDl which is over expressed in majority of the cancers was identified as a direct target of miR 17-5p (8). Under expression status of miR 125 a & b in HER 2 positive tumours indicated their role as a tumour suppressor of this gene.
  • MicroRNAs miR 21 , miR155 and miRlOb have been shown to play a role in tumour metastasis by targeting anti metastatic genes (12, 13, 14).
  • MiR-21 is over expressed in both male and female breast tumors compared with normal breast tissue and has been associated with advanced stage, lymph node positivity, and reduced survival time.
  • existence of microRNAs either floating or in exosomes in the systemic circulation has led to the possibility that such molecules may serve as biomarkers for early detection of cancers.
  • microRNA profiling is emerging as a powerful tool for diagnosis of breast cancer types, grades and stages.
  • the main objective of the present invention relates to biomarkers for diagnosis of different types, grades and stages of human breast cancer.
  • Another objective of the present invention relates to molecular biomarkers as indicators of cellular changes during the initiation and development of breast cancer.
  • Yet another objective of the present invention relates to a chip useful for detection and diagnosis of breast cancer.
  • Still another objective of the present invention is to provide a cheaper, accurate, robust and high throughput diagnostic kit for accurate diagnosis of human breast cancer.
  • the present invention relates to a panel of Biomarkers useful for screening and detection for the type, grade and stage of Breast cancer wherein the panel comprises of miRNA having sequence selected from the group consisting of Seq ld No. 1- 107.
  • a panel of Biomarkers useful for screening and detection for the type, grade and stage of Breast cancer wherein the panel comprises of miRNA having sequence selected from the group consisting of Seq Id No. 1 - 107
  • the panel of Biomarkers wherein down regulation of microRNAs with Seq ID No. 1 to 12 and up regulation of Seq ID No. 13 to 15 detects ER +ve type of breast cancer.
  • the panel of Biomarkers wherein down regulation of Seq.ID no. 16 & 17 and up regulation of Seq. ID no. 18 to 29 detects ER- Ve type of breast cancer.
  • the panel of Biomarkers wherein down regulation of Seq.ID no. 30 to 36 and up regulation of Seq.ID no. 37 to 42 detects grade 2 breast cancer.
  • the panel of Biomarkers wherein down regulation of Seq.ID no. 43 to 48 and up regulation of Seq.ID no. 49 & 50 detects grade 3 breast cancer.
  • the panel of Biomarkers wherein down regulation of Seq.ID no. 56 to 73 detects stage II of grade 2 breast cancer.
  • the panel of Biomarkers wherein down regulation of Seq.ID no. 74 & 75 and up regulation of Seq.ID no. 76 to 81 detects stage III of grade 2 breast cancer.
  • the panel of Biomarkers wherein down regulation of Seq.ID no. 82 to 95 and up regulation of Seq.ID no. 96 & 97 detects stage I of grade 3 breast cancer.
  • the panel of Biomarkers wherein down regulation of Seq.ID no. 98 to 100 and up regulation of Seq.ID no. 101 to 103 detects stage II of grade 3 breast cancer.
  • the panel of Biomarkers wherein down regulation of Seq.ID no. 104 & 105 and upregulation of Seq.ID no. 106 & 107 detects stage III of grade 3 breast cancer.
  • the panel of Biomarkers wherein the antisense sequences of the upregulated & downregulated microRNAs is of therapeutic use.
  • a panel of miRNA in the form of a DNA/ RNA chip in yet another embodiment of the present invention a panel of miRNA in the form of a DNA/ RNA chip
  • kits for detecting type, grade and stage of breast cancer consisting of :
  • Suitable reagents capable of detecting singly or a combination of the miRNA Instruction manual for using the kit.
  • kit Suitable reagents capable of detecting singly or a combination of the miRNA; Instruction manual for using the kit.
  • biomarkers for detection of type, grades and stages of Breast Cancer.
  • kits for detection of type, grades and stages of Breast Cancer use of the kit for detection of type, grades and stages of Breast Cancer.
  • Table 1 The sequence IDs 1 to 15 lists the microRNAs which are significantly up/down regulated in ER +ve human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are lip regulated indicated by -Ve sign) are also given.
  • Table 2 The sequence IDs 16 to 29 lists the microRNAs which are significantly up/down regulated in ER -ve human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated indicated by -Ve sign) are also given.
  • Table 3 The sequence IDs 30 to 42 lists the microRNAs which are significantly up/down regulated in grade 2 human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated 9indicated by -Ve sign) are also given.
  • Table 4 The sequence IDs 43 to 50 lists the microRNAs which are significantly up/down regulated in grade 3 human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated indicated by -Ve sign) are also given.
  • Table 5 The sequence IDs 51 to 55 lists the microRNAs which are significantly up/down regulated in stage I of grade 2 human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated indicated by -Ve sign) are also given.
  • Table 6 The sequence IDs 56 to 73 lists the microRNAs which are significantly up/down regulated in stage II of grade 2 human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated Syndicated by -Ve sign) are also given.
  • Table 7 The sequence IDs 74 to 81 lists the microRNAs which are significantly up/down regulated in stage III of grade 2 human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated 9indicated by -Ve sign) are also given.
  • Table 8 The sequence IDs 82 to 97 lists the microRNAs which are significantly up/down regulated in stage I of grade 3 human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated 9indicated by -Ve sign) are also given.
  • Table 9 The sequence IDs 98 to 103 lists the microRNAs which are significantly up/down regulated in stage II of grade 3 human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated 9indicated by -Ve sign) are also given.
  • Table 10 The sequence IDs 104 to 107 lists the microRNAs which are significantly up/down regulated in stage III of grade 3 human breast cancer samples.
  • the microRNA names along with their sequences and accession IDs are also described here.
  • the exact fold change with which each microRNA is down regulated (indicated by +ve sign) and those which are upregulated 9indicated by -Ve sign) are also given.
  • Table 11 rniRNAs validated and reconfirmed by Individual Taqman assays in different grades and stages of Breast cancer spotted on a biochip.
  • Fig 1 The sequence IDs 1 to 15 lists the microRNAs which are significantly up/down regulated in ER +ve and non-significant in ER -ve human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 2 The sequence IDs 16 to 29 lists the microRNAs which are significantly up/down regulated in ER -ve and non-significant in ER +ve human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 3 The sequence IDs 30 to 42 lists the microRNAs which are significantly up/down regulated in grade 2 and non-significant in grade 3 human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 4 The sequence IDs 43 to 50 lists the microRNAs which are significantly up/down regulated in grade 3 and non-significant in grade 2 human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 5 The sequence IDs 51 to 55 lists the microRNAs which are significantly up/down regulated in Stage I of grade 2 and non-significant in stage II and III of grade 2 human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 6 The sequence IDs 56 to 73 lists the microRNAs which are significantly up/down regulated in Stage II of grade 2 and non-significant in stage I and III of grade 2 human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 7 The sequence IDs 74 to 81 lists the microRNAs which are significantly up/down regulated in Stage III of grade 2 and non-significant in stage I and II of grade 2 human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 8 The sequence IDs 82 to 97 lists the microRNAs which are significantly up/down regulated in Stage I of grade 3 and non-significant in stage II and III of grade 3 human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 9 The sequence IDs 98 to 103 lists the microRNAs which are significantly up/down regulated in Stage II of grade 3 and non-significant in stage I and III of grade 3 human breast cancer samples.
  • the heat map represents the up and down regulation with respective p values.
  • Fig 10 The sequence IDs 104 to 107 lists the microRNAs which are significantly tip/down regulated in Stage III of grade 3 and non-significant in stage I and II of grade 3 human breast cancer samples.
  • the heat map represents the up and down regulation with, respective p values.
  • Breast cancer is a complex heterogeneous genetic disease, involving a variety of changes in gene expression and structure.
  • MicroRNAs are recently discovered tiny RNA molecules which play an important role in the gene regulation. They are found to have an altered expression in majority of cancers and are termed as oncomiRs.
  • oncomiRs Recently, advances in molecular profiling has shed new light on the etiology of the disease and also marketed great potential for the development of novel biomarkers for diagnosis, prognosis and therapeutic targets. This attracts the scientific domain for extensive investigation to further elucidate their precise role as novel biomarkers in malignancy.
  • MicroRNAs are tiny biological molecules that play a regulatory role in biological processes and cellular functions. Therefore these molecules could be used as indicators of changes in the cells, when they transform from normal to diseased condition.
  • This invention specifically relates to the identification of changes in these small RNA regulations that play an important role in the development of breast cancer.
  • oncomiRs an expression signature of these microRNAs involved in cancer
  • oncomiRs changes in the expression pattern of small RNAs called oncomiRs from breast cancer patients at different grades and stages of development of cancer.
  • the expression profile of these miRNAs formed a classic signature, as breast cancer progressed from stage I to stage III, in both, grades.
  • These differentially up and down regulated microRNAs are significant in one type, stage and grade of cancer and not in the other. Therefore we have classified them into type, grade and stage specific biomarkers which could be useful tools in the diagnosis and prognosis of breast cancer
  • miRNAs possess unique features that classify them as ideal tumor markers include their tissue specificity, stability, ease of detection and association with the disease status.
  • miRNAs have vast possibility in diagnosis, prognosis and treatment of diseases especially malignancies like breast cancer, where still no reliable tumor markers for particular stages and grades are available at present.
  • these molecular biomarkers can be used to accurately identify subgroups of patients who will really benefit from cytotoxic chemotherapy with its debilitating side effects. Thus this proves to be an additional, accurate, quick and high throughput molecular diagnostic tool for pathologists especially when patient number is high.
  • miRNAs are ideal and potential biomarkers for detecting different grades and stages of breast cancer. A few to hundred samples can be checked within a span of 2 to 3 hrs and hence this becomes an easy, fast and high throughput technology.
  • microRNAs are present in the cells and altered signatures are detected and reported in cancer samples, the identification of the different subtype, grade and stage specific microRNAs along with its fold regulation demarcated them as ideal biomarkers for breast cancer diagnosis, prognosis and targeted therapy.
  • microRNAs are identified by LNA microarray and is verified with Taqman Low Density arrays. The fold regulation of each microRNA was also found by TLDA analysis. Additional validation of these microRNAs were carried out using Taqman individual assays in individual cancer samples samples and identified for making this as a diagnostic chip (Table 1 1).
  • Megaplex Reverse Transcription rxn (40 cycles) is done using Megaplex RT Primers, (TaqMan® MicroRNA Reverse Transcription Kit) -dNTPs with dTTP, MultiscribeTM reverse Transcriptase, 10 X RT Buffer and RNase Inhibitor.
  • the reverse transcription (RT) reaction was done in a final volume of 7.5 ⁇ , which contains: 3 ⁇ , total RNA and 4.5 of RT reaction mix.
  • Thermocycling condition was set as default and Ramp speed or mode: 9700 using Std or Max ramp.
  • pre amplified specific cDNA targets were subjected to increase, the quantity of desired cDNA for miRNA expression analysis using TaqMan® MicroRNA Arrays.
  • the preamplification reaction was performed in a final volume of 25 ⁇ ⁇ _ where: 2.5 ⁇ RT product and 22.5 ⁇ PreAmp reaction mix was present.
  • Threshold cycles were compared and analyzed using arithmetic formulas that determines the change in expression of a target gene in an experimental sample relative to the same target in a reference sample. This method was used for high- throughput measurements of relative gene expression.
  • Statminer software was used for fold expression analysis of miRNAs and classified as detector not amplified, significant and nonsignificant based on their p values. Highly significant (>0.05) miRNAs were selected for study and biomarker identification in the respective types of breast cancer.
  • Example 1 Total RNA isolation and quality control
  • RNA Integrity Number RNA Integrity Number
  • MicroRNA profiling was done using TaqManMicroRNA Arrays, which contains megaplex Primer Pools covering Sanger miRBase versionl O.
  • Megaplex Reverse Transcription Primers are novel stem-looped RT primer pools that streamline the profiling of hundreds of miRNA targets in a single experiment and reduce the number of Reverse Transcription reactions and the amount of total RNA required for generating a comprehensive miRNA expression profile.
  • a pre amplification step of cDNA with preamp megaplex pool primers was done to significantly enhance the ability to detect slowly expressed miRNAs.
  • Example 3 Real Time amplification of miRNA pool by loading in TLDA plates
  • the TaqMan human MicroRNA arrays consists of 2 plates pool A and pool B. 'A' Array Sanger's V 10.0 contains 667human taqman microRNA Assays. Three TaqMan MicroRNA Assay. Endogenous controls are included for data normalization and one TaqMan MicroRNA Assay, not related to human is also included as a negative control. The set enables accurate quantitation of 667 human microRNAs. Results of Taqman Low Density Arrays analysis
  • statminer software This contains a filtering procedure for outlier removal; various normalization methods based on single or multiple genes and provides relative quantification analysis of gene 5 expression through a combination of statistical analysis and interactive visualization.
  • CT threshold cycle
  • microRNAs were selected which pertains to different subtypes (ER+ve, ER-ve), grades and stages (Table 1, 2 and 3).
  • RNA isolated from the same cancer samples were hybridized against 2002 microRNAs consisting of 10 904 human, 388 rat and 710 mouse microRNAs. The normals were labeled with Hy5
  • the normalized median signal intensities for the Hy3 (sample) and Hy5 (common reference) indicate the relative expression level of each microRNA in the samples and 15 in the common reference. If the Hy3 value is higher than the Hy5 value there is a
  • Hy3 higher expression in the sample than in the common reference and if the Hy3 value is lower than that of Hy5 value there is a lower expression in the samples compared to the common reference. Then we take the ratios between the Hy3 and Hy5 signal and the log2 to that ratio. A positive number indicates a higher expression in the sample (Hy3) compared to the common reference and vice versa.
  • the NA means that the microRNA is not expressed based on certain ctit-off criteria. One criterion is the signal intensity of the Hy3 and Hy5 channel. If both Hy3 and Hy5 signals are below 1.5 times the median of all capture probes on the array we say that it is background and below our cut-off. This cut-off is set to avoid too many false positives.
  • Call rate is the number of expressed microRNAs compared to the total number of microRNAs analyzed (the % of identified microRNAs). This call rate is expected to be between 20 and 50% for human samples and it is clear that we have a very nice and high call rate in our samples. That human samples have call rates between 20 and 50% has been documented in the literature, both based on deep sequencing, array and PCR profiling. A call rate much higher than 50% indicates a high risk of having false positives in the data set Therefore we used the 1.5X median of all capture probes as a cut off.
  • microRNAs have been analyzed based on the samples groups. A two-tailed statistical t-test has been performed between the samples groups grade 2 and grade 3. The heat map has been made based on a cut-off of P ⁇ 10 "3 . "Expression matrix (analysis)” looked like typical breast cancer microRNAs. A very long list of breast cancer miRNAs from literature, web databases are all present in our samples. Just to mention some examples, miR-21, miR-155, miR-148a, miR210 and miR-29b.These typical breast cancer signatures clearly classify our samples as breast cancer samples.
  • miRNAs in particular stages or grades shows its behavior which is highly correlated with the expression of translational regulators or targets that are involved in tumor progression.
  • the miRNAs which are down regulated at stage I gets up or down regulated successively at stage II and stage III with in a grade. This classical pattern of miRNA expression indicates their importance in controlling the progressive growth of breast cancer.
  • biomarkers could be developed as a diagnostic kit for early and accurate diagnosis of human breast cancer. They are direct indicators of cellular changes during the initiation and development of breast cancer. These biomarkers complement the pathologists for the accurate grading and staging of breast cancer. These biomarkers provided a utility angle to the already existing biological molecules called microRNAs which play a major role in gene regulation. These biomarkers could provide a fast, cheaper, accurate, robust and high throughput diagnostic kit for accurate diagnosis of human breast cancer.
  • Table 11 miRNAs validated and reconfirmed by Individual Taqman assays in different grades and stages of Breast cancer spotted on a biochip.
  • micro-ribonucleic acid (miRNA) miR-206 targets the human estrogen receptor-alpha (ERalpha) and represses ERalpha messenger RNA and protein expression in breast cancer cell lines. Mol Endocrinol 2007, 21 : 1 132-1 147.
  • MicroRNA- 155 is regulated by the transforming growth factor beta/Smad pathway and contributes to epithelial cell plasticity by targeting RhoA. Mol Cell ⁇ ⁇ /2008, 28:6773-6784.

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Abstract

L'invention concerne des biomarqueurs servant à détecter des types, des niveaux et des stades du cancer du sein chez l'être humain. Cette invention concerne en particulier le développement desdits biomarqueurs identifiés en tant que puce à microARN pour un diagnostic précoce et précis du cancer du sein chez l'être humain. La nouveauté de cette invention réside dans l'utilité de ces microARN, dans la mesure où il peuvent être employés comme kit de diagnostic (puce à micro-ARN) pour réaliser une détection précoce et précise de niveaux, stades et sous-types du cancer du sein. Quelques échantillons à plusieurs centaines d'échantillons peuvent être vérifiés en l'espace de 2 à 3 heures, ce qui permet d'obtenir une technologie simple, rapide, robuste et à haut rendement pour un programme de criblage à des fins de détection précoce du cancer du sein.
PCT/IB2012/002090 2011-10-19 2012-10-17 Biomarqueurs servant à détecter des types, niveaux et stades du cancer du sein chez l'être humain WO2013057567A1 (fr)

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CA2852384A CA2852384C (fr) 2011-10-19 2012-10-17 Biomarqueurs servant a detecter des types, niveaux et stades du cancer du sein chez l'etre humain
US14/352,804 US10208353B2 (en) 2011-10-19 2012-10-17 Biomarkers useful for detection of types, grades and stages of human breast cancer
AU2012324531A AU2012324531B2 (en) 2011-10-19 2012-10-17 Biomarkers useful for detection of types, grades and stages of human breast cancer
GB1408696.1A GB2509682B (en) 2011-10-19 2012-10-17 Biomarkers useful for detection of types, grades and stages of human breast cancer
AU2018202963A AU2018202963B2 (en) 2011-10-19 2018-04-30 Biomarkers useful for detection of types, grades and stages of human breast cancer
US16/219,572 US11136628B2 (en) 2011-10-19 2018-12-13 Biomarkers useful for detection of types, grades and stages of human breast cancer

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IN1142/DEL/2011 2011-10-19
IN1142DE2011 2011-10-19

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US16/219,572 Division US11136628B2 (en) 2011-10-19 2018-12-13 Biomarkers useful for detection of types, grades and stages of human breast cancer

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US10208353B2 (en) 2011-10-19 2019-02-19 Council Of Scientific And Industrial Research Biomarkers useful for detection of types, grades and stages of human breast cancer
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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10208353B2 (en) 2011-10-19 2019-02-19 Council Of Scientific And Industrial Research Biomarkers useful for detection of types, grades and stages of human breast cancer
US11136628B2 (en) 2011-10-19 2021-10-05 Council Of Scientific And Industrial Research Biomarkers useful for detection of types, grades and stages of human breast cancer
WO2015136141A1 (fr) * 2014-03-13 2015-09-17 Universidad De Málaga Signature de micro-arn utile comme indicateur du risque de récidive précoce chez des patientes atteintes du cancer du sein
US10059998B2 (en) 2014-03-13 2018-08-28 Servicio Andaluz De Salud Microrna signature as an indicator of the risk of early recurrence in patients with breast cancer
ES2481819A1 (es) * 2014-06-12 2014-07-31 Sistemas Genómicos, S.L. Método de evaluación para evaluar una posibilidad de cáncer de mama
WO2015189444A1 (fr) * 2014-06-12 2015-12-17 Sistemas Genómicos, S.L. Méthode pour évaluer l'éventualité d'un cancer du sein
EP3268494A4 (fr) * 2015-03-09 2018-11-07 Agency For Science, Technology And Research Procédé de détermination du risque de développer un cancer du sein par détection des niveaux d'expression de micro-arn (miarn)
EP3070178A1 (fr) * 2015-03-20 2016-09-21 Albert-Ludwigs-Universität Freiburg Procédé de diagnostic du cancer du sein
US10246749B2 (en) 2015-03-20 2019-04-02 Albert-Ludwigs-Universitaet Freiburg Method for diagnosing breast cancer
CN108070654A (zh) * 2017-04-13 2018-05-25 朱伟 肺癌miRNA检测的内参基因的鉴定
CN108070654B (zh) * 2017-04-13 2019-03-01 朱伟 肺癌miRNA检测的内参基因的鉴定
CN111154878A (zh) * 2020-02-14 2020-05-15 北京粒基生物科技有限公司 一组miRNA在制备作为用于早期乳腺癌筛查的标志物中的应用

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GB201603127D0 (en) 2016-04-06
CA3077746A1 (fr) 2013-04-25
CA3077800A1 (fr) 2013-04-25
AU2012324531A1 (en) 2014-05-15
US20150080244A1 (en) 2015-03-19
GB2536374B (en) 2016-10-26
GB2535914B (en) 2016-10-26
GB2533872A (en) 2016-07-06
AU2018202963A1 (en) 2018-05-17
GB2533872B (en) 2016-08-31
GB2535914A (en) 2016-08-31
US11136628B2 (en) 2021-10-05
GB2509682B (en) 2016-07-27
GB201608844D0 (en) 2016-07-06
GB2509682A (en) 2014-07-09
US20190106754A1 (en) 2019-04-11
GB2533873B (en) 2016-08-31
GB201608839D0 (en) 2016-07-06
GB2536374A (en) 2016-09-14
CA2852384C (fr) 2020-06-23
CA3077750A1 (fr) 2013-04-25
GB2533873A (en) 2016-07-06
GB201603128D0 (en) 2016-04-06
CA3077750C (fr) 2022-05-10
AU2012324531B2 (en) 2018-03-22
US10208353B2 (en) 2019-02-19
GB201408696D0 (en) 2014-07-02
AU2018202963B2 (en) 2020-11-26

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